<?xml version="1.0" encoding="ISO-8859-1"?><article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance">
<front>
<journal-meta>
<journal-id>2221-2450</journal-id>
<journal-title><![CDATA[Revista CENIC Ciencias Biológicas]]></journal-title>
<abbrev-journal-title><![CDATA[Rev. CENIC Cienc. Biol]]></abbrev-journal-title>
<issn>2221-2450</issn>
<publisher>
<publisher-name><![CDATA[Centro Nacional de Investigaciones Científicas]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S2221-24502021000100011</article-id>
<title-group>
<article-title xml:lang="en"><![CDATA[Production of a human papillomavirus type 18-His-tag-L1 protein by a working cell bank of Escherichia coli BL21 (DE-3)]]></article-title>
<article-title xml:lang="es"><![CDATA[Producción de una proteína del virus del papiloma humano tipo 18-His-tag-L1 por un banco de células de trabajo de Escherichia coli BL21 (DE-3)]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Serrano-Díaz]]></surname>
<given-names><![CDATA[Anabel]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Pimienta-Rodríguez]]></surname>
<given-names><![CDATA[Elsa]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Brito-Molina]]></surname>
<given-names><![CDATA[Susana]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Serrano-Rivero]]></surname>
<given-names><![CDATA[Yunier]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Falero-Morejón]]></surname>
<given-names><![CDATA[Alina]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[la O-Reyes]]></surname>
<given-names><![CDATA[Rosa A.]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Marrero-Domínguez]]></surname>
<given-names><![CDATA[Karen]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,National Center for Scientific Research Development &amp; Innovation Research Unit of Biological Products, Division of Research]]></institution>
<addr-line><![CDATA[Playa La Habana]]></addr-line>
<country>Cuba</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>04</month>
<year>2021</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>04</month>
<year>2021</year>
</pub-date>
<volume>52</volume>
<numero>1</numero>
<fpage>11</fpage>
<lpage>17</lpage>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_arttext&amp;pid=S2221-24502021000100011&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_abstract&amp;pid=S2221-24502021000100011&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_pdf&amp;pid=S2221-24502021000100011&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="en"><p><![CDATA[ABSTRACT  The infection with Human Papilloma viruses (HPV) is the most common sexually transmitted disease worldwide. The HPV 16 and 18 types are responsible for around 70% of cervical cancer cases globally. In Cuba, this is the second cancer in incidence and mortality on women between 15 to 44 years old. The viral capsid is formed by 72 capsomeres, composed by L1 and L2 proteins, being L1 the major protein. The available preventive vaccines are based on virus-like particles (VLPs) assembled from L1 protein produced in eukaryotic hosts and they have high prices, which has hindered their deployment in middle- and low-income countries. The bacterium Escherichia coli has been used as an alternative cheaper host for second-generation HPV vaccine production. The aims of this study were to prepare a working cell bank (WCB) of E. coli BL21 (DE3) harboring the HPV18-His-tag-L1-pET28a plasmid, which encodes the HPV18-L1 protein fused to a six-histidine-Tag; as well as to evaluate L1 protein´s production and solubility under isopropyl-&#946;-D-1-thiogalactopyranoside (IPTG) induction and on auto-inducing conditions, as the first step to tackle L1 purification. The highest L1 levels were detected on terrific broth (TB)-grown cultures induced for 4h or 16h with low-IPTG-concentrations (30 and 60 µmolL-1) or with a lactose/glucose mixture for 16 hours. In both conditions, the protein was mainly detected in the insoluble fraction, so its purification will be carried out under denaturing conditions. The available WCB will allow maintaining the constant quality and homogeneity of the initial inoculum during L1 production to develop a robust purification process.]]></p></abstract>
<abstract abstract-type="short" xml:lang="es"><p><![CDATA[RESUMEN La infección con el virus del Papiloma Humano (VPH) es la enfermedad de transmisión sexual más común en el mundo. El VPH16 y el 18 son responsables de cerca del 70% de los casos de cáncer de cuello uterino a nivel global. En Cuba, este es el segundo tipo de cáncer, tanto en incidencia como en mortalidad, en mujeres en edades comprendidas entre los 15 y 44 años. La cápside del virus está formada por 72 capsómeros, compuestos por las proteínas L1 y L2, siendo L1 la proteína mayoritaria. Las vacunas preventivas disponibles se basan en partículas semejantes a virus ensambladas a partir de L1 producida en hospederos eucariotas y presentan elevados precios, lo que ha limitado su uso en los países de ingresos medios y bajos. La bacteria Escherichia coli se ha empleado como un hospedero alternativo y más barato para la producción de vacunas de segunda generación contra el VPH. Los objetivos de este estudio fueron preparar un banco de células de trabajo de E. coli BL21 (DE3) con el plasmidio HPV18-His-Tag-L1-pET28a, que codifica para la proteína L1 de VPH18 fusionada a una cola de seis Histidinas (His); así como evaluar la producción y solubilidad de dicha proteína en condiciones de inducción con isopropil-&#946;-D-1-tiogalactopiranósido (IPTG) y de autoinducción, como primer paso para abordar la purificación de la proteína L1. Los mayores niveles de la proteína se detectaron en los cultivos en medio terrific broth (TB) inducidos con bajas concentraciones de IPTG, 30 y 60 µmol/L, y en los que se usó una mezcla de lactosa/glucosa por 16h. En ambos casos, la proteína se encontró mayoritariamente en la fracción insoluble, de modo que su purificación deberá realizarse en condiciones desnaturalizantes. El banco de células de trabajo preparado permitirá mantener constante la calidad y homogeneidad del inóculo inicial durante la producción de L1, lo que posibilitará desarrollar un proceso de purificación robusto.]]></p></abstract>
<kwd-group>
<kwd lng="en"><![CDATA[human papillomavirus]]></kwd>
<kwd lng="en"><![CDATA[Escherichia coli]]></kwd>
<kwd lng="en"><![CDATA[L1 protein]]></kwd>
<kwd lng="en"><![CDATA[HPV18]]></kwd>
<kwd lng="es"><![CDATA[virus del papiloma humano]]></kwd>
<kwd lng="es"><![CDATA[Escherichia coli]]></kwd>
<kwd lng="es"><![CDATA[proteína L1]]></kwd>
<kwd lng="es"><![CDATA[VPH18]]></kwd>
</kwd-group>
</article-meta>
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